正畸静压力通过p38MAPK调控牙周膜细胞MMP-2和MMP-9的表达

Static Orthodontic Pressure Regulates the Expression of MMP-2 and MMP-9 in Periodontal Ligament Cells via p38MAPK Pathway

目的:研究持续性正畸静压力对牙周膜细胞(PDLCs)表达MMP-2和MMP-9的调控及其有关的调控通路。方法:体外培养PDLCs,取4~7代的PDLCs,随机分成5组,分别施加0、2 g/cm^(2)(24 h)、2 g/cm^(2)(48 h)、4 g/cm^(2)(24 h)、4 g/cm^(2)(48 h)的持续静压力,对细胞形态及骨架F-actin进行检测,通过Western blot观察持续静压力对MMP-2、MMP-8、MMP-9的表达的影响。研究相关MAPK通路包括JNK、ERK、p38MAPK在静压力引起MMPs表达变化中的作用。分组添加p38MAPK阻断剂SB203580,分别通过Western blot和免疫荧光观察p38MAPK通路的作用。结果:在不同的持续静压力下,PDLCs形态未见明显变化,F-actin染色显示细胞骨架未有明显区别。Western blot显示静压力对MMP-8表达未见明显影响,MMP-2和MMP-9的表达明显上调。且这种上调作用与p38MAPK通路有关,与JNK和ERK通路无关。结论:持续静压力调控PDLCs MMP-2和MMP-9的表达,这种调控通过p38MAPK途径产生作用,与JNK和ERK通路无关。

Objective:To instigate the regulation effects of static pressure on the expression of MMP-2 and MMP-9 in periodontal ligament cells(PDLCs)and to explore related pathways.Methods:PDLCs were cultured in vitro.Cells of 4-7 generations were randomly divided into 5 groups,i.e.0,2 g/cm^(2)(24 h),2 g/cm^(2)(48 h),4 g/cm^(2)(24 h),and 4 g/cm^(2)(48 h)groups.Continuous static pressure was used to detect cell morphology and skeleton F-actin.The effect of continuous static pressure on the expression of MMP-2,MMP-8,and MMP-9 was observed by western blot.Related MAPK pathways including JNK,ERK,and p38MAPK in the static pressure induced expression of MMPs were studied by western blotting.p38MAPK blocker SB203580 was added into each groups,and the effect of p38MAPK was observed by western blot and immunofluorescence.Results:Under different continuous static pressures,there was no obvious change in the morphology of PCLCs.F-actin staining showed no significant difference in cytoskeleton.Western blotting showed that static pressure had no significant effect on the expression of MMP-8,and the expressions of MMP-2 and MMP-9 were significantly up-regulated.This upregulation was related to the p38MAPK pathway,but not to the JNK and ERK pathways.Conclusion:Continuous static pressure regulates the expression of MMP-2 and MMP-9 in periodontal ligament cells.This regulation is related to the p38MAPK pathway but JNK and ERK pathways.